Invitrogen

ARF6 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

4 Published Figures

7 References

View all (14) ARF6 antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

ARF6 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of ARF6 was performed using 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with ARF6 Polyclonal Antibody (Product # PA1-093) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification.

ARF6 Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on normal biopsies of deparaffinized human heart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a Rabbit Polyclonal Antibody recognizing Arf6 (Product # PA1-093) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

ARF6 Antibody in Western Blot (WB) — Western Blot was performed using Anti-ARF6 Polyclonal Antibody (Product # PA1-093) and a 25 kDa band corresponding to ARF6 was observed across cell lines with uncharacterized band(*). Whole cell extracts (30 µg lysate) of LNCaP (Lane 1), MDA-MB-231 (Lane 2), HUVEC (Lane 3), MCF7 (Lane 4), MCF 10A (Lane 5), SK-BR-3 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody ( 1 mg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

ARF6 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. MDA-MB-231 cells were transfected with ARF6 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-ARF6 Polyclonal Antibody (Product # PA1-093). {KD}

ARF6 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Figure 3 Secretion of sFlt1 from endothelial cells requires functional Arf6, Arf1, and Rab11 GTPases. A-F : EC-sFlt1 cells were nucleofected with plasmid constructs expressing dominant-negative forms of Arf1, Arf6, Rab5, Rab6, Rab 8, syntaxin 4, syntaxin 6, and syntaxin 16. Similarly, in a separate study EC-sFlt1 cells were nucleofected with scramble oligonucleotide or siRNAs against Arf1 , Arf6 , and Rab11a . Media containing nucleofection complexes were replaced with fresh media 8 hrs post-nucleofection. At 36 hrs post-nucleofection, secreted and intracellular sFlt1 were immunoprecipitated from spent culture media and cell lysates using the anti-v5 Ab. ( A , C ) Representative blot shows relative expression of sFlt1-v5 in cell lysates and secreted in culture media. B , D : Band densities from experiments as in A, C were quantified by densitometric analysis. Secretion of sFlt1-v5 was calculated as the ratio of desitometric band intensity values of secreted to intracellular Flt1; this ratio was set at 1 for mock-transfected control EC-sFlt1 cells. The data are the means +- SEM from three independent experiments. E : EC-sFlt1 cells were transiently transfected with indicated dominant-negative plasmid constructs. After 24 hrs cells were fixed, permeabilized, and labeled with anti-v5 Ab to detect cell-associated sFlt1-v5. Representative images obtained by epifluorescence microscopy show localization of sFlt1-v5. F : Quantification of intracellular retention of sFlt1-v5 upon inhi

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1,000

View 4 publications 4 publications

Immunohistochemistry (Paraffin) (IHC (P))

1:20-100

Immunocytochemistry (ICC/IF)

1:20-1:100

View 2 publications 2 publications

Miscellaneous PubMed (Misc)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Human, Mouse, Non-human primate

Published species

Human, Mouse

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

C-terminal human recombinant protein fragment

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

PBS with 1mg/mL BSA, 30% glycerol

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2539860

Target Information

ARF6 (ADP-ribosylation factor 6) is a broadly expressed, differentially regulated GTPase that is present in cytoplasm and on both cell-surface and intracellular membranes and whose functions may include tissue-specific effects on vesicular trafficking during cellular differentiation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.